posted on 2020-03-12, 13:10authored byMontserrat Mancera-Arteu, Estela Giménez, Jaime Sancho, Victoria Sanz-Nebot
Transferrin purification
from mice serum samples by immunoaffinity
chromatography (IAC) was optimized in order to study the possible
modifications occurring in its glycans in collagen-induced arthritis
(CIA) samples. SDS-PAGE and nanoLC-MS/MS were used to monitor the
IAC purification performance. Afterward, a relative quantification
of mouse transferrin (mTf) glycan isomers using [12C6]/[13C6]-aniline was used to unequivocally
detect alterations in the glycan profile of CIA mice. In addition,
multivariate data analysis was applied to identify the most meaningful
glycan isomers for the discrimination between control and pathological
samples. Partial least-squares discriminant analysis (PLS-DA) revealed
that five out of fifteen mTf glycan isomers could be potential biomarkers
of CIA, most of them corresponding to highly sialylated structures
(H6N5S3_2, H6N5S3_3, and H5N4S3_2). Moreover, some of these glycan
isomers also seemed to be related with the progression of CIA, especially
H6N5S2 and H6N5S3_2, as their overexpression increased with the clinical
score of the pathology. Hence, the established methodology not only
provides valuable information to find glycan-based biomarkers of CIA,
but also leaves the door open to evaluate, in the future, glycosylation
changes of many other inflammatory diseases, in which transferrin
has been described to be altered.